Recent in silico searching of the human genome yielded three related proteins - the pannexins (Panxl, Panx2 and Panx3), originally speculated to function as gap junction channels because of the their connexin- like protein domain organization. Although connexins are the most widely studied intercellular communication channels, non-connexin gap junction channels have been discovered experimentally in invertebrates (innexins) and pannexins are more similar to innexins than connexins. The tissue structure analyses proposed in this research will investigate, using large-scale fluorescence light microscopic montage imaging, Panxl and Panx2 expression in the rodent brain to determine areas with high levels of expression and then use data sharing, mining and neuroinformatic tools to see whether pannexins co- localize with other proteins as interaction partners or indicate that they are part of a larger cellular complex. This project aims to integrate experimental imaging with computational analyses in order to obtain quantitative and interdisciplinary information about pannexin expression/function. This proposal utilizes a data-intensive mosaic imaging technique for large-scale mapping of the mouse brain combined with neuroinformatics, data mining, and sharing to help integrate and synthesize data for the broader impact of modeling the complex mammalian brain. Specifically, the goals of my research are: (1) To apply our recently developed antibody tool kit to image whole mouse brain at high light microscopic resolution where both Panxl and Panx2 are highly expressed, (2) To deposit and annotate the brain maps in public databases for data sharing, (3) To use the brain maps in conjugation with neuroinformatics and data mining computational tools to determine if there are "hot spots" for Panxl and Panx2 localization in the mouse brain correlated with high expression of other proteins such as connexins or purinergic receptors, (4) To correlate membrane versus intracellular localizations of endogenous Panxl and Panx2 in tissue to complement ex vivo studies of exogenously expressed pannexins in cell culture. Relevance: This is part of a multi-scale coordinated approach with directions in molecular, cellular and organ level imaging of Panxl and Panx2. This multilevel approach addresses the particular challenge for nervous system research to bridge the dimensional range from tissues to molecules, a range encompassing cellular networks, dendritic and axonal architectures, synaptic connectivity, glial interactions and macromolecular constituents. These structures, and the proteins within them, represent the heart of information processing in the nervous system;the integration, synthesis, and sharing of this protein expression data is central to our understanding of brain function.